Review: Recent advances in insect-based feeds: from animal farming to the acceptance of consumers and stakeholders.

The search for new, alternative and sustainable feeding sources, including insects, has become an important challenge on the feed market. In 2017, the European Union (EU) started to allow the use of insect meals as feeds for fish. In addition, in 2021, the EU also authorised the use of insect meal for pig and poultry farming. However, the adoption of insect meal by the European aquaculture sector is still limited, and this is mostly due to the lack of availability of insects and their higher costs than conventional feed ingredients. Thus, the insect-based feed industry is still in its infancy, and its successful development and integration in the food value chain depend on several factors. Among these, the technical feasibility and production of quality products, and acceptance by European consumers and farmers are relevant factors. To address these points, this narrative review describes the state of the art of the potential role of insect-based feeds. The stakeholders' and consumers' perspectives are investigated, along with the effects of insect-based feeds on the production and nutritional values of fish, poultry (meat and eggs), and pork. Indeed, matching the nutritional values of insect products with conventional feeds is one of the future challenges of the insect sector, as their nutritional composition is highly dependent on the rearing substrates, and thus, their use in animal feeding needs to be investigated carefully. Feeding animals with insect-based diets affects their growth performances and the chemical composition of the derived products (fish fillets, meat, and eggs). Whether these effects can be considered positive or negative seems to depend to a great extent on the percentage of insects included in their diets and the chemical composition of the ingredients. The use of insect-based feeds has also shown a potential to improve the nutritional features and values of animal products and even to add new ones. Finally, many of the acceptance studies on the use of insects in feeds have focused mostly on the consumers' perception rather than on industry stakeholders (e.g., farmers). Future research should focus more on the farmers' perceptions on and market analyses of these innovative feeds. Even though it is likely that the upscaling of the insect sector will lead to a decrease in prices and an increase in market availability, it is still critical to understand the potential barriers and drivers for the implementation of insects as feeds from a production point of view.

Determination of lipid requirements in black soldier fly through semi-purified diets.

The insect market is still far from an effective upscale and, to achieve this goal, it is necessary to know the BSF dietary requirements for the production maximization. Worldwide, given the waste variability, is not always easy to identify the optimal waste-based mixture that can allow to reach the best production, in terms of quantity and quality. Due this reason, nutritional need ranges are the basic knowledge, affordable for everyone, to increase the profitability of the insect farming. The study aims to evaluate the effects of 6 semi-purified, isonitrogenous and isoenergetic diets (SPII) with increasing lipid levels (1%, L1; 1.5%, L1.5; 2.5%, L2.5; 3.5% L3.5; 4.5%, L4.5) on BSF life history traits (6 replicates/treatment and 100 larvae/replicate). The Gainesville diet was used as environmental control. Considering the whole larval stage, 4.5% lipid level guarantees better performance when compared to content lower than 2.5%. The L4.5 10-day-old larvae yielded greater when compared to the other dietary treatments. At 14 and 18 days of age, the larvae of the groups above 2.5% performed better than L1, while the L1.5 showed intermediate results. Lipid levels below 1.5% on DM, when compared to 4.5%, resulted in a smaller prepupa and pupa size. The results obtained on the adult stage do not allow the identification of a lipid levels ideal range, as in the larval stage. In conclusion, in the whole larval stage and in prepupae/pupae phases, lipid percentage lower than (or equal to) 1% have a negative effect on growth. Other research will be needed in order to evaluate lipid levels above 4.5% on DM.

Black soldier fly larva in Muscovy duck diets: effects on duck growth, carcass property, and meat quality.

The aim of the study was to evaluate the effects of partially defatted black soldier fly (Hermetia illucens, HI) larva meal on the carcass characteristics and meat quality of Muscovy ducks (Cairina moschata domestica). A total of 192 female ducks aged 3 d were divided between 4 dietary treatments (6 pens/treatment; 8 birds/pen), characterized by increasing levels of substitution of corn gluten meal with HI meal (0%, 3%, 6%, and 9%; HI0, HI3, HI6, and HI9, respectively), and reared until 50 days of age. Twelve birds/treatment (2 birds/pen) were slaughtered on d 51 to evaluate the slaughter traits (i.e., carcass, breast, thigh, and organs weights), carcass yield and meat quality. The slaughter weight, hot and chilled carcass weights, and abdominal fat weight showed a quadratic response to HI meal (minimum for the HI6 group, P < 0.05). Dietary HI meal inclusion did not influence the ultimate pH, the color, the proximate composition or the thiobarbituric acid reactive substances (TBARS) values in either the breast or thigh meat. The mineral profile of the meat was slightly affected by the dietary treatment, with a linear increase in the Cu content of the thigh meat (P < 0.05), whereas no differences were observed for Zn, Mn, or Fe. Dietary HI meal inclusion increased the saturated fatty acid rate in the thigh meat (maximum for the HI9 group, P < 0.05), and the monounsaturated and polyunsaturated fatty acid content in the breast meat (maximum for the HI0 and HI9 groups, respectively, P < 0.05). The ∑n-6/∑n-3 ratio decreased linearly in both the breast and thigh meat, with the HI9 group showing the lowest values (P < 0.05). Finally, the heavy metal concentrations were below the EU limits for poultry meat. To conclude, the inclusion up to 9% of partially defatted HI larva meal in the diet of Muscovy ducks did not affect the slaughter traits or the meat quality, although it did affect the meat fatty acid profile.

Black soldier fly defatted meal as a dietary protein source for broiler chickens: effects on carcass traits, breast meat quality and safety.

Finding insect meals as alternative sources of poultry feedstuffs is a recent research topic; therefore, the present study aimed to evaluate the effects of defatted black soldier fly (Hermetia illucens L., HI) larvae meal in broiler chicken diets on the carcass characteristics and meat quality parameters, proximate composition, fatty acid profile and the heavy metal content of the breast meat. Four dietary treatments were designed: a control diet (HI0) and three experimental diets (HI5, HI10 and HI15), corresponding to 50, 100 and 150 g/kg HI inclusion levels, respectively. The inclusion of 50, 100 and 150 g HI meal per kg feed supply 16.56%, 33.01% and 49.63% of required crude protein. The broilers were slaughtered at day 35, the carcasses were weighed and the breast muscles were excised from 16 birds per each feeding group (two birds per replicate pens) and used for meat quality evaluation. Linear and quadratic responses were observed, for increasing HI meal levels, in the live and carcass weights (maximum for HI10). As far as the colour of the breast meat is concerned, redness (a*) showed a linear response, while yellowness (b*) linearly decreased with increasing HI meal levels (minimum for HI15). As the HI larvae meal increased in the diets, the moisture content linearly decreased and the protein content increased. The total saturated fatty acid and total monounsaturated fatty acid proportions rose to the detriment of the polyunsaturated fatty acid fraction. The HI larvae meal, used in the current study, represents a valuable protein source for broiler chickens when included by up to 100 g/kg in their diets, as an improved slaughtering performance was observed without any detrimental effects on meat quality parameters or heavy metal residues in the meat.

Bilberry pomace in rabbit nutrition: effects on growth performance, apparent digestibility, caecal traits, bacterial community and antioxidant status.

Agricultural by-products could be used as alternative raw materials in rabbit nutrition as they have been found to be highly nutritious and low cost feeding sources. The aim of this study was to estimate the nutritive value and potential use of bilberry pomace (BP) for growing rabbits. A total of 144 Grimaud rabbits (35 days old) were allotted to four groups and fed with a diet containing increasing level of BP: BP0 (basal diet), BP5, BP10 and BP15 containing 0, 50, 100 and 150 g/kg respectively. Growth trial lasted 48 days; apparent digestibility was evaluated, starting at 46 days of age, over 4 consecutive days. The nutritive value of BP was measured using the mean digestibility of the experimental diets. At 83 days of age, rabbits were slaughtered: blood, and liver and kidney samples were collected in order to determine the blood parameters and the antioxidant enzyme activities of the tissues. Moreover, caecal content was sampled and gut microbiota assessed by means of amplicon-based high-throughput 16S rRNA sequencing and PCR-denaturing gradient gel electrophoresis. The digestible protein was estimated to 104 g/kg of DM while digestible energy to 9.44 MJ/kg DM for incorporation rate up to 150 g/kg. During the finishing period, average daily feed intake and feed conversion ratio showed linear response to BP increase (P=0.008 and <0.001, respectively). During all the period, both parameters decreased linearly and quadratically with increasing BP inclusion levels (P<0.001) up to 100 g/kg of BP. A significant effect of the antioxidant status was found in the kidneys and liver (P<0.05) where the glutathione peroxidase activity increased as the BP increased. As far as gut microbiota is concerned, BP increased the relative abundance of the Clostridium, Oscillospira, Ruminococcus and Ruminococcaceae species which were clearly associated with the BP inclusion level. In conclusion, BP showed a potential use as an alternative protein and fibre sources for growing rabbits.

Effect of partial dietary replacement of fishmeal by yellow mealworm (Tenebrio molitor) larvae meal on the innate immune response and intestinal antioxidant enzymes of rainbow trout (Oncorhynchus mykiss).

The present study investigated the effects of 25 and 50% dietary inclusion of yellow mealworm, Tenebrio molitor (TM), larvae on the intestinal antioxidant enzymes, lipid peroxidation and immune system of rainbow trout, Oncorhynchus mykiss. Fish were fed for 90 days with a control diet (TM0) or with 2 test diets (TM25 and TM50) where 35 and 67% of the fishmeal was replaced by TM larvae meal respectively. At the end of the growth trial, humoral immune parameters were assessed in the sera of the fish and the activities of antioxidant enzymes were measured in intestine homogenates. The present results show a strong increase of the activity of the antioxidant intestinal enzymes and reduction of lipid peroxidation. Increased trypsin inhibition and faster antibacterial activity of the serum were also detected. These evidences could be due to the physiological similarities between the exoskeleton of parasites and insects. The present findings reinforce the previous results which revealed a positive effect of the dietary TM on rainbow trout growth and further confirm its adequacy for fishmeal partial replacement in aquafeed.

Quality of eggs from Lohmann Brown Classic laying hens fed black soldier fly meal as substitute for soya bean.

Soya bean is the main protein source in poultry feed but rising prices make an alternative protein source necessary. Insects, such as the black soldier fly (Hermetia illucens), may be an attractive solution for hens, although little information is available on their effect on egg quality. The present study aims to fill this gap by testing the effect of 100% replacement of soya bean with H. illucens larva meal in the diet of Lohmann Brown Classic laying hens for 21 weeks. At the end of the trial, the eggs were characterized for parameters such as weight, colour, proximate composition of albumen and yolk, and content of carotenoids, tocopherols and cholesterol. The fatty acid profile of yolks was also determined. Hens fed the insect-based diet produced eggs (HIM group) with a higher proportion of yolk than the group fed the soya bean-based diet (SBM group). HIM was associated with redder yolks (red index 5.63 v. 1.36) than SBM. HIM yolks were richer in γ-tocopherol (4.0 against 2.4 mg/kg), lutein (8.6 against 4.9 mg/kg), ß-carotene (0.33 against 0.19 mg/kg) and total carotenoids (15 against 10.5 mg/kg) than SBM yolks. The fatty acid composition of HIM yolks was almost identical to that of SBM yolks. Finally, HIM yolks contained 11% less cholesterol than SBM yolks. These results suggest that H. illucens larva meal is a suitable total substitute for soya bean meal in the diet of Lohmann Brown Classic laying hens. A sustainable alternative to the plant protein source therefore seems feasible.

Black soldier fly larva fat inclusion in finisher broiler chicken diet as an alternative fat source.

The objective of the present study was to evaluate the effects of partial or total replacement of finisher diet soybean oil with black soldier fly (Hermethia illucens L.; HI) larva fat on the growth performance, carcass traits, blood parameters, intestinal morphology and histological features of broiler chickens. At 21 days of age, a total of 120 male broiler chickens (Ross 308) were randomly allocated to three experimental groups (five replicates and eight birds/pen). To a basal control diet (C; 68.7 g/kg as fed of soybean oil), either 50% or 100% of the soybean oil was replaced with HI larva fat (HI50 and HI100 group, respectively). Growth performance was evaluated throughout the trial. At day 48, 15 birds (three birds/pen) per group were slaughtered at a commercial abattoir. Carcass yield and proportions of carcass elements were recorded. Blood samples were taken from each slaughtered chicken for haematochemical index determination. Morphometric analyses were performed on the duodenum, jejunum and ileum. Samples of liver, spleen, thymus, bursa of fabricius, kidney and heart were submitted to histological investigations. Growth performance, carcass traits, haematochemical parameters and gut morphometric indexes were not influenced by the dietary inclusion of HI larva fat. Histopathological alterations developed in the spleen, thymus, bursa of fabricius and liver and were identified in all of the experimental groups, but HI larva fat inclusion did not significantly affect (P>0.05) the severity of the histopathological findings. The present study suggests that 50% or 100% replacement of soybean oil with HI larva fat in broiler chickens diets has no adverse effects on growth performance or blood parameters and had no beneficial effect on gut health.

Yellow mealworm larvae (Tenebrio molitor) inclusion in diets for male broiler chickens: effects on growth performance, gut morphology, and histological findings.

This study evaluated the effects of Tenebrio molitor (TM) larvae meal inclusion in diets for broilers. A total of 160 male broiler chicks (Ross 708) at one-day of age were randomly allotted to four dietary treatments: a control (C) group and three TM groups, in which TM meal was included at 50 (TM5), 100 (TM10), and 150 (TM15) g/kg, respectively. The experimental diets were isonitrogenous and isoenergetic. Each group consisted of five pens as replicates (8 chicks/pen). After the evaluation of growth performance and haematochemical parameters, the animals were slaughtered at 53 days and carcass traits were recorded. Morphometric investigations were performed on duodenum, jejunum, and ileum and histopathological alterations were assessed for liver, spleen, thymus, bursa of Fabricius, kidney, and heart. The live weight (LW) showed a linear (12 and 25 days, P < 0.001 and P < 0.05, maximum with TM15 and TM10) and quadratic (53 days, P < 0.05, maximum with TM5) response to dietary TM meal inclusion. A linear (1 to 12 and 12 to 25 days, P < 0.001, maximum with TM15) and quadratic (12 to 25 days, P = 0.001, maximum with TM15) effect was also observed for the daily feed intake (DFI). The feed conversion ratio (FCR) showed a linear response (25 to 53 and 1 to 53 days, P = 0.001 and P < 0.05, maximum with TM15). Haematological and serum biochemical traits, carcass traits and histopathological findings were not affected by dietary TM meal inclusion (P > 0.05). TM15 birds showed lower villus height (P < 0.05), higher crypt depth (P < 0.05), and lower villus height to crypt depth ratio (P = 0.001) compared with C and TM5. In conclusion, increasing levels of dietary TM meal inclusion in male broiler chickens may improve body weight and feed intake, but negatively affect feed efficiency and intestinal morphology, thus suggesting that low levels may be more suitable. However, no effect on haematochemical parameters, carcass traits, and histological findings were observed in relation to TM meal utilization.

Does dietary insect meal affect the fish immune system? The case of mealworm, Tenebrio molitor on European sea bass, Dicentrarchus labrax.

Feeding small European sea bass, Dicentrarchus labrax, for 6 weeks with Tenebrio molitor larval meal showed significant anti-inflammatory responses (ceruloplasmin, myeloperoxidase and nitric oxide). Serum bacteriolytic activity against a Gram negative bacterium was not significantly affected by dietary Tenebrio, while both lysozyme antibacterial activity and serum trypsin inhibition usually linked to the anti-parasite activity of the fish, were significantly enhanced. The latter may be due to the similarities in the composition of the exoskeleton of parasites and insects that may therefore act as an immunostimulant potentially increasing the anti-parasitic activity. The addition of exogenous proteases significantly decreased both trypsin-inhibition and serum bacteriolytic activity probably through direct inhibition of the proteins responsible for these immune functions. Further investigation involving bacterial or parasitic challenges will be necessary to assess if the effects of dietary mealworm meal on the immune system observed in the present study are translated into an improved resistance to diseases.

Laser treatment of post-thyroidectomy scar.

A good aesthetic result in thyroid surgery is one of the main goals, as this procedure usually involves young women affected in clearly visible anatomical areas. To minimize the detrimental effects of scarring outcome, several therapeutic options have been employed. Lasers may be an alternative choice for prevention and treatment of post-surgical thyroidectomy scar. This paper reviews literature and the current knowledge on this topic. A comprehensive search in the Cochrane Library, MEDLINE and PUBMED databases was performed to identify relevant literature investigating the role of laser therapy in both prevention and treatment of unappealing scarring after thyroid surgery. Laser treatment of post-thyroidectomy scar is emerging with promising clinical outcomes. The greatest efficacy has been seen with vascular-selective and ablative sources. Laser therapy should be taken into account as it represents a valid and safe treatment option.

Evaluation of the suitability of a partially defatted black soldier fly (Hermetia illucens L.) larvae meal as ingredient for rainbow trout (Oncorhynchus mykiss Walbaum) diets.

BACKGROUND: Two trials were performed to evaluate a partially defatted Hermetia illucens (HI) larvae meal as potential feed ingredient in rainbow trout (Oncorhynchus mykiss Walbaum) diets. In the first trial, 360 trout (178.9 ± 9.8 g of mean initial body weight) were randomly divided into three experimental groups (4 tanks/treatment, 30 fish/tank). The fish were fed for 78 days with isonitrogenous, isolipidic and isoenergetic diets containing increasing levels of HI, on as fed basis: 0% (HI0, control diet), 25% (HI25) and 50% (HI50) of fish meal substitution, corresponding to dietary inclusion levels of 0, 20% and 40%. In the second trial, 36 trout (4 tanks/treatment, 3 fish/tank) were used to evaluate the in vivo apparent digestibility coefficients (ADC) of the same diets used in the first trial. RESULTS: Survival, growth performance, condition factor, somatic indexes, and dorsal fillet physical quality parameters were not affected by diet. The highest dietary inclusion of HI larvae meal increased dry matter and ether extract contents of trout dorsal fillet. The use of HI larvae meal induced a decrease of valuable polyunsaturated fatty acids (PUFA) even if differences were only reported at the highest level of HI inclusion. The insect meal worsened the lipids health indexes of the same muscle. Dietary inclusion of insect meal did not alter the villus height of the fish. No differences were found among treatments in relation to ADC of ether extract and gross energy, while ADC of dry matter and crude protein were higher in HI25 if compared to HI50. CONCLUSIONS: The obtained results showed that a partially defatted HI larvae meal can be used as feed ingredient in trout diets up to 40% of inclusion level without impacting survival, growth performance, condition factor, somatic indexes, dorsal fillet physical quality parameters, and intestinal morphology of the fish. However, further investigations on specific feeding strategies and diet formulations are needed to limit the observed negative effects of the insect meal on the FA composition of dorsal muscle.

Productive performance and blood profiles of laying hens fed Hermetia illucens larvae meal as total replacement of soybean meal from 24 to 45 weeks of age.

The aim of the research was to study the effects of an insect meal from Hermetia illucens larvae (HILM) as complete replacement of soybean meal (SBM) on productive performance and blood profiles of laying hens, from 24 to 45 wk of age. A total of 108 24-week-old Lohmann Brown Classic laying hens was equally divided into 2 groups (54 hens/group, 9 replicates of 6 hens/group). From 24 to 45 wk of age, the groups were fed 2 different isoproteic and isoenergetic diets: the control group (SBM) was fed a corn-soybean meal based diet, while in the HILM group the soybean meal was completely replaced by Hermetia illucens larvae meal. Feed intake, number of eggs produced, and egg weight were recorded weekly along the trial. At 45 wk of age, blood samples were collected from 2 hens per replicate. The use of HIML led to a more favorable (P < 0.01) feed conversion ratio in hens but lay percentage, feed intake, average egg weight, and egg mass were higher (P < 0.01) in hens fed the SBM diet. Hens fed insect meal produced a higher percentage of eggs from small (S), medium (M), and extra-large (XL) classes (P < 0.01) than SBM, while the SBM group had a higher percentage of eggs from the large (L) class (P < 0.01). The levels of globulin and albumin to globulin ratio were, respectively, higher and lower (P < 0.05) in HILM than the SBM group. Cholesterol and triglycerides were higher (P < 0.05 and P < 0.01, respectively) in hens from SBM than in the HILM group. Blood levels of Ca were higher (P < 0.01) in hens fed insect meal, while creatinine was higher (P < 0.01) in blood of hens fed SBM. Hermetia illucens larvae meal can be a suitable alternative protein source for laying hens even if the complete replacement of soybean meal needs further investigation to avoid the negative effects on feed intake.

Use of larvae meal as protein source in broiler diet: Effect on growth performance, nutrient digestibility, and carcass and meat traits.

The aim of this research was to study the effect of insect meal from larvae ( larvae meal [TML]) as complete replacement of soybean meal (SBM) on growth performance, nutrient digestibility, and carcass and meat traits of broilers. A total of eighty 30-d-old male Shaver brown broilers were homogenously divided into 2 groups (each consisting of 8 replicates of 5 birds). Up to 62 d of age, the groups were fed 2 isoproteic and isoenergetic diets differing for the ingredient used as the main protein source: the control group was fed a corn-SBM-based diet, whereas in the TML group, the SBM was completely replaced by TML. Broiler growth performance was measured during the trial. At 62 d of age, 2 broilers per replicate (16 per group) were slaughtered and apparent ileal digestibility coefficients and carcass and meat traits were determined. The use of TML as the main protein source in the broiler diet had no significant effect on most growth performance and carcass traits and chemical and physical properties of meat, the latter being important for marketing purposes. The feed conversion ratio in the entire experimental period (from 30 to 62 d) was improved in the TML group compared with the SBM group ( < 0.05). The apparent ileal digestibility coefficients of DM, OM, and CP in broilers fed the SBM diet were greater ( < 0.01) than the other group. The full digestive system in broilers fed SBM had a lower ( < 0.05) absolute and relative weight than that of broilers fed TML. Also, the weight and the percentage of the spleen in the SBM group were lower ( < 0.05) than those in the TML group. The length of the entire intestine in the group fed TML was greater ( < 0.05) than the other group and the same happened when intestinal length was expressed as percentage of broiler BW ( < 0.05). Among the different intestinal tracts, the ileum and ceca of broilers fed TML had a greater ( < 0.05) length than that of broilers fed SBM. Also, ceca weight (as an absolute value or percentage on live weight) in broilers fed TML was greater ( < 0.05) than that of broilers fed SBM. Breast pH and cooking losses in broilers fed TML were greater ( < 0.05) than those of broilers fed SBM. Chemical characteristic of meat obtained from right breast of slaughtered broilers were unaffected by dietary treatments. larvae meal can be a suitable alternative protein source for growing broilers and also when used as principal protein contributor to the diet.

Oxidative stress related to chlorpyrifos exposure in rainbow trout: Acute and medium term effects on genetic biomarkers.

Organophosphates (OPs) are derivatives of phosphoric acid widely used in agriculture as pesticides. Chlorpyrifos (CPF) is an OP that is extremely toxic to aquatic organisms. Rainbow trout (Oncorhynchus mykiss) is considered as a sentinel model species for ecotoxicology assessment in freshwater ecosystems. An exposure study was carried out on rainbow trout to investigate genetic responses to CPF-induced oxidative stress by Real-Time PCR, and to determine the accumulation dynamics of CPF and toxic metabolite chlorpyrifos-oxon (CPF-ox) in edible parts, by HPLC-MS/MS. Among the genes considered to be related to oxidative stress, a significant increase in HSP70 mRNA levels was observed in liver samples up to 14 days after CPF exposure (0.05 mg/L). CPF concentrations in muscle samples reach mean values of 285.25 ng/g within 96 hours of exposure, while CPF-ox concentrations were always under the limit of quantification (LOQ) of the applied method. Our findings lead us to consider HSP70 as a suitable genetic marker in rainbow trout for acute and medium-term monitoring of CPF exposure, complementary to analytical determinations.

Effects of dietary Tenebrio molitor meal inclusion in free-range chickens.

Insects are currently being considered as a novel protein source for animal feeds, because they contain a large amount of protein. The larvae of Tenebrio molitor (TM) have been shown to be an acceptable protein source for broiler chickens in terms of growth performance, but till now, no data on histological or intestinal morphometric features have been reported. This study has had the aim of evaluating the effects of dietary TM inclusion on the performance, welfare, intestinal morphology and histological features of free-range chickens. A total of 140 medium-growing hybrid female chickens were free-range reared and randomly allotted to two dietary treatments: (i) a control group and (ii) a TM group, in which TM meal was included at 75 g/kg. Each group consisted of five pens as replicates, with 14 chicks per pen. Growth performance, haematological and serum parameters and welfare indicators were evaluated, and the animals were slaughtered at the age of 97 days. Two birds per pen (10 birds/treatment) were submitted to histological (liver, spleen, thymus, bursa of Fabricius, kidney, heart, glandular stomach and gut) and morphometric (duodenum, jejunum and ileum) investigations. The inclusion of TM did not affect the growth performance, haematological or serum parameters. The morphometric and histological features were not significantly affected either, thus suggesting no influence on nutrient metabolization, performance or animal health. Glandular stomach alterations (chronic flogosis with epithelial squamous metaplasia) were considered paraphysiological in relation to free-range farming. The observed chronic intestinal flogosis, with concomitant activation of the lymphoid tissue, was probably due to previous parasitic infections, which are very frequently detected in free-range chickens. In conclusion, the findings of this study show that yellow mealworm inclusion does not affect the welfare, productive performances or morphological features of free-range chickens, thus confirming that TM can be used safely in poultry diets.

Rabbit dietary supplementation with pale purple coneflower. 1. Effects on the reproductive performance and immune parameters of does.

Echinacea pallida (EPAL), also known as pale purple coneflower, is a herbaceous flowering plant with immune-enhancement and antioxidative properties. The effect of EPAL on the reproductive performance, serum biochemistry and haematological parameters of rabbit does has been studied here. A total of 100, 21-week-old Grimaud rabbit does, were randomly assigned to two groups. One group was fed a basal diet supplemented with 3 g EPAL/kg diet (Echinacea group, E), while the other was fed the basal diet without the supplementation (control group, C). The reproductive performance of the does was not affected by the treatment (P>0.05). The haematological parameters of pregnant rabbits showed that there was no interaction between gestation day and treatment. The EPAL supplementation induced a reduction (-47.3%) in the basophil cell rate (0.55% and 0.29%, for the control and treatment groups, respectively; P=0.049). The gestation day significantly affected most of the haematological parameters (P<0.05). The white blood cell counts declined progressively after day 14. The mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration, red cell distribution width, mean platelet volume and eosinophils increased steadily throughout the study, and reached a maximum value on day 28. The red blood cells, haemoglobin, haematocrit, mean corpuscular volume and neutrophils increased slightly up to day 14, and then subsequently decreased progressively until day 28. The lymphocytes and platelet distribution width decreased until day 14, and then increased to a maximum value on day 28. No significant effect of gestation day or treatment was observed on the blood serum chemistry. As far as the immune parameters are concerned, no significant differences were observed between groups, while a significant effect of gestation day was observed for lysozymes (6.02 v. 7.99 v. 1.91; for 0, 14 and 28 days, respectively; P=0.014). In conclusion, a lack of effect of EPAL has been observed. In fact, no impacts of EPAL have been observed on the reproductive or haematological parameters of the does. The effects of dietary supplementation with EPAL on the performances, bacterial community, blood parameters and immunity in growing rabbits are reported in the second part of this study.

Rabbit dietary supplementation with pale purple coneflower. 2. Effects on the performances, bacterial community, blood parameters and immunity of growing rabbits.

Echinacea pallida (EPAL), a herbaceous flowering plant with immunomodulatory properties, has been chosen to determine the pre- and post-supplementary effects on the growth performances, bacterial community, blood parameters and immunity of growing rabbits. The same Grimaud does (14-week-old) from the studied in the first part of this study were randomly divided into two groups (n=50/group). The first group was fed a basal diet without supplementation (Control group, C) while the another group was fed a basal diet supplemented with 3 g EPAL/kg diet (Echinacea group, E). From the second parturition, 80 weaned kits (40 from the C does and 40 from the E does) were randomly assigned to four groups of 20 animals each and were fed a growing commercial diet supplemented with or without a 3 g EPAL/kg diet: the CC group (rabbits from the C does fed the control diet), CE group (rabbits from the C does fed the supplemented diet), EC (rabbits from the E does fed the control diet) and EE group (rabbits from the E does fed the supplemented diet). The dietary EPAL treatment did not affect the growth performance. Ten fattening rabbits from each group were selected to evaluate the bacterial community and blood parameters, while the remaining rabbits (n=10/group) were used to study phagocytosis and the humoral immune response. The variability was evaluated from hard faeces at 35, 49 and 89 days, and the caecal content at 89 days. The variability of the bacterial community of the EE group was higher than that of the other groups. The phagocytic activity was higher in the CE and EE groups than in the CC and EC ones (30.9 and 29.7 v. 21.2 and 21.8%; P<0.05), whereas no statistically significant difference was observed for the blood parameters or humoral immune response against vaccination (rabbit haemorrhagic disease virus) at 95 days old which the serum was collected at 88, 102, 109, 116 and 123 days old. In conclusion, no impact of EPAL dietary supplementation has been observed on the growth performances, bacterial community, blood parameters or humoral immune responses in growing rabbits, except for an increase in phagocytic activities.

Effect of purple loosestrife (Lythrum salicaria) diet supplementation in rabbit nutrition on performance, digestibility, health and meat quality.

In this study, 160 Hycole weaned rabbits (35 days old) were randomly divided into four groups of 40. The rabbits were studied throughout a 54-day experimentation period in order to determine the impact of dietary supplementation from herbs composed of 0.2%, 0.4% dry ground Lythrum salicaria leaves (LS) and 0.3% Cunirel(®) (CR; a commercial herb mixture containing LS as the main ingredient) on performance, digestibility, health and meat quality. The basal diet was given to the control group. No significant differences were found in performance, 10 rabbits from each group were selected for evaluation regarding apparent digestibility. The rabbits fed the control diet and the diet with the low level of LS had a higher level of CP digestibility than did the animals that were supplemented with the high LS levels and CR (85.7% and 84.9% v. 84.0% and 84.0%, respectively; P<0.05). The ether extract digestibility was lower in the treatment group with 0.4%LS addition and CR as compared with the control group (52.2% and 54.5% v. 62.6%, respectively; P<0.05). The slaughter process was performed on 89-day-old rabbits to study the carcass characteristics, meat quality, blood parameters, caecal contents and gut histology. The total leukocyte counts in the control animals were lower than they were in the rabbits fed 0.2%, 0.4%LS and CR (4.06 v. 8.25, 8.63 and 8.21×10(9)/l, respectively; P<0.05). For caecal fermentation, the caecal contents of the rabbits fed 0.4% of LS, showed higher concentrations of total volatile fatty acid (VFA; 24.1 v. 18.9 mg/kg dry matter (DM); P<0.05) and acetic acid (18.3 v. 14.4 mg/kg DM; P<0.05), but lower ammonia levels (594 v. 892 mg/kg DM; P<0.05) as compared with the control group. PCR-denaturing gradient gel electrophoresis analyses were performed to evaluate the microbial community in hard faeces, collected at days 35, 42, 49, 56, 70 and 89, whereas the caecal contents were taken after slaughtering. The results demonstrated that between the treatment groups, the similarity of the microbial communities was higher as compared with the control group. Moreover, only age was shown to influence microbiota diversity. In conclusion, the results of this study indicated that supplementation of LS in rabbit diets leads to an increase in the total white blood cells, total VFA and acetic acid concentration, and a decrease in the ammonia levels, as well as the digestibility when CR and high level of LS were supplemented, without causing any adverse effects on other parameters.